Study On The Differentiation Of Mesenchymal Stem Cells Into Cardiomyocyte-like Cells By H9C2Cell Culture Medium

Objective: To investigate the role of H9C2cell culture medium in thedifferentiation of mesenchymal stem cells（MSCs）into cardiomyocyte-likecells。Method: MSCs were obtained by whole bone marrow adherent cultureand H9C2cell culture medium was prepared as a culture medium,thenMSCs were co-cultured with H9C2cell culture medium for1、3、5、7days.H9C2cells which express specific markers of myocardial cells as positivecontrol group, MSCs induced by H9C2cell culture medium to differentiateas the experimental group, MSCs untreated as negative control group;Detect each group’s myocardial cell characteristics after the proliferationand differentiation of MSCs cells. Immunofluorescence and western-blotdetect expression of myocardial cell junction protein (desmin) and troponinT (cTnT). Real-time quantitative PCR (RT-PCR) detect mRNA ’s expressionof myocardial cell trait gene α-cardiac myosin heavy chain (a-MHC) and theexpression of β-myosin heavy chain (β-MHC). Result:1H9C2cell culture medium co-culture with MSCs for7days,Immunofluorescence detect expression of troponin T of cells in each group,the cTnT positive cells during MSCs were up to0.179±0.011, which wassignificantly higher (P <0.05)than non-induced MSCs.2H9C2cell culture medium co-culture with MSCs for7days,western-blot detect the expression of troponin T, desmin of cells in eachgroup, the expression of MSCs’s cTnT protein and desmin after inductionwere significantly increased (P <0.05) by western-blot detection;3H9C2cell culture medium co-culture with MSCs for7days,RT-PCR detect the expression of α-MHC and β-MHC mRNA, RT-PCRshowed that differentiated cells mRNA expression of α-MHC and β-MHCwere up-regulated (P <0.05).Conclusion: H9C2cell culture medium may induce the differentiationof MSCs into cardiomyocyte-like cells.